Method for treating late phase allergic reactions and inflammatory diseases

ABSTRACT

A method of treating a mammalian patient suffering from or prone to a condition characterized by late phase allergic reactions, airway hyperresponsiveness or inflammatory reactions, e.g., asthma, allergic rhinitis, allergic dermatitis, allergic conjunctivitis, inflammatory bowel disease or rheumatoid arthritis, comprising the administration to the patient of an oral, parenteral, intrabronchial, topical, intranasal or intraocular pharmaceutical composition containing in each dose about 0.005 to about 1.0 mg per kilogram of patient body weight of ultra-low molecular weight heparins (ULMWH) or other sulfated polysaccharides having average molecular weights of about 1,000-3,000 daltons. Suitable inhalant and other pharmaceutical compositions for use in the novel treatment method are also disclosed.

REFERENCE TO DISCLOSURE DOCUMENT

This application incorporates material included in Disclosure DocumentNo. 401115, filed in the Patent and Trademark Office on Jun. 5, 1996.

CROSS-REFERENCE TO RELATED APPLICATION

This application is a continuation-in-part of application Ser. No.08/516,786, filed Aug. 18, 1995, now U.S. Pat. No. 5,690,910 issued Nov.15, 1997.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The invention relates to methods and compositions for preventing andreversing the symptoms and manifestations of late phase allergicreactions and inflammatory diseases.

2. Description of the Prior Art

Chronic asthma can be considered to be predominantly an inflammatorydisease with associated bronchospasm. The degree of reactivity andnarrowing of the bronchi in response to stimuli is greater in asthmaticsthan in normal individuals. Persistent inflammation is responsible forthe bronchial hyperreactivity or airway hyperresponsiveness (AHR).Mucosal edema, mucus plugging and hypersecretion may be present;pulmonary parenchyma is normal. Airway narrowing may reversespontaneously or with therapy. Type 1 (immediate) immune responses mayplay an important role in the development of asthma in children and manyadults; however, when onset of disease occurs in adulthood, allergicfactors may be difficult to identify. Exposure to cold dry air, exerciseand other aggravating factors also may trigger asthma.

The general goals of drug therapy for asthma are prevention ofbronchospasm and long-term control of bronchial hyperreactivity. Becauseit is usually not possible for either patient or physician to predictwhen bronchospasm may occur, patients with all but the most episodicand/or entirely seasonal attacks may require continuous therapy.

Beta agonists are useful as bronchodilator agents; they stimulate beta₂-adrenergic receptors, increase intracellular cAMP, and may inhibit therelease of mast cell mediators. Other useful drugs include theophyllineand related xanthine drugs, which produce bronchodilation throughunknown mechanisms; the biscromone, cromolyn, which prevents the releaseof mediator substances and blocks respiratory neuronal reflexes; andcorticosteroids, which primarily decrease inflammation and edema.Anticholinergic drugs may relieve bronchospasm by blockingparasympathetic cholinergic impulses at the receptor level.Antihistamines occasionally prevent or abort allergic asthmaticepisodes, particularly in children, but they can only be partiallyeffective in asthma because histamine is only one of many mediators.

The current drug modalities used for treatment of allergy-induced asthmasuffer from a number of drawbacks. In general, the conventional agentshave a relatively short duration of action and may be partially orwholly ineffective when administered after antigen challenge occurs.Moreover, because of serious adverse effects associated with the use ofagents such as beta₂ -adrenergic agonists and corticosteroids, thetherapeutic margin of safety with such agents is relatively narrow andpatients using them must be carefully monitored.

Bronchial hyperreactivity (or AHR) is a hallmark of asthma and isclosely related to underlying airway inflammation. Worsening of asthmaand airway inflammation is associated with increase in bronchialhyperreactivity, which can be induced by both antigenic andnon-antigenic stimuli. Beta₂ -adrenergic agonists are potent agents forthe treatment of bronchospasm, but have no effect on airway inflammationor bronchial hyperreactivity. In fact, chronic use of beta₂ -adrenergicagents alone, by causing down regulation of beta₂ -receptors, may worsenbronchial hyperreactivity. At present, corticosteroids are the onlyeffective agents available which diminish bronchial hyperreactivity.Although inhaled corticosteroids are relatively safe in adult patientswith asthma, these agents have tremendous toxicity in children,including adrenal suppression and reduced bone density and growth. Thus,the search for safer and effective agents which diminish bronchialhyperreactivity continues.

Patients with allergic asthma, following an inhalation challenge withthe specific antigen exhibit at least two different patterns ofbronchial responses. The majority of subjects develop an acutebronchoconstrictor response only, which resolves spontaneously within1-3 hours; these subjects are termed "acute responders". A smallernumber of subjects, however, develop both an early and a late response.These subjects are termed "dual responders". In dual responders, theacute response, which resolves spontaneously, is followed 4-12 hourslater by a secondary increase in airway resistance ("late phaseresponse"). Late responses and thus dual responders are of clinicalimportance, because of their association with prolonged airwayhyperreactivity or hyperresponsiveness (AHR), worsening of symptoms andgenerally worse form of clinical asthma, requiring aggressive therapy.

Pharmacological studies in allergic animals have demonstrated that notonly the bronchoconstrictor response but also the inflammatory cellinflux and the mediator release pattern in dual responders is quitedifferent from acute responders. Whereas histamine is the likelybronchoconstrictor mediator during acute phase, activated products ofthe lipoxygenase pathway (i.e., leukotrienes) may be the major mediatorinvolved in late phase reaction. Mast cells, however, have a centralrole in IgE-mediated allergic airway responses, and cromolyn sodium (amast-cell membrane stabilizer), theoretically should preventbronchoconstrictor responses in both "acute" and "dual" responders.Heterogeneity of mast cell subtypes may play a significant role indivergent responses and it may be dependent upon differences in signaltransduction (second messenger system).

It has been discovered in the past several years that heparinadministered intrabronchially can be an effective inhibitor ofbronchospasm and bronchoconstriction and is consequently of value in theprophylaxis of asthma (see, e.g., Ahmed et al., New Eng. J. Med.,329:90-95,1993; Ahmed, Resp. Drug Deliv., IV:55-63, 1994). It has beendiscovered further that low molecular weight heparins, e.g., heparinswith an average molecular weight of 4,000-5,000 daltons, effectivelyprevent antigen-induced bronchoconstriction; these low molecular weightheparins also exhibit considerably less anticoagulant activity thancommercial heparin, a desirable property when these agents are used inthe treatment of asthma (see Ashkin et al., Am. Rev. Resp. Dis., 1993Intl. Conf. Abstracts, p. A660). Both commercial and low-weight heparinsare not effective, however, in suppressing AHR when administered afterthe patient has been exposed to antigen.

In parent application Ser. No. 08/516,786 we disclosed that ultra-lowmolecular weight heparins (ULMWH) having an average molecular weightless than about 3,000 daltons are effective in suppressing AHR in acuteasthmatic responders, even when administered after the patient has beenchallenged with antigen. However, experimental and clinical studies haveshown that while inhaled commercial heparin can also attenuate earlyphase antigen-induced bronchoconstriction in acute responders (thoughnot after antigen challenge) it is ineffective in the treatment of dualresponders. Hence, there was still considerable doubt after our earlierwork with ULMWH as to whether these substances would show efficacy inthe treatment of dual or late responders as they do in acute responders.

The current, conventional therapeutic modalities for asthmatic patientswho are dual responders are generally a more aggressive andtime-prolonged version of the therapies practiced on acute responders,described above. However, these therapies are not particularly effectivein suppressing AHR, as noted previously, and, as a result of theirgenerally short duration of action, cannot prevent the late phasereaction and AHR observed in dual responders.

It should be noted, however, that the airways are merely a prototype oforgans or tissues affected by late phase reactions (LPR's). It has beenestablished in the medical literature that the late phasebronchoconstriction and AHR observed in dual responder asthmaticpatients is not an isolated phenomenon restricted to asthmatic or evenpulmonary conditions. There are cutaneous, nasal, ocular and systemicmanifestations of LPR's in addition to the pulmonary ones. Theseallergic LPR phenomena are considered closely interrelated from thepoint of view of immunologic mechanisms. See Lemanske and Kaliner, "LatePhase Allergic Reactions", published in Allergies. Principles andPractice (Mosby Yearbook, Inc., 4th ed. 1997). According to the latestunderstanding of LPR mechanisms it appears that the clinical diseases(whether of the skin, lung, nose, eye or other organs) recognized toinvolve allergic mechanisms have a histologic inflammatory componentwhich follows the immediate allergic or hypersensitivity reaction thatoccurs on antigen challenge. This sequence of response appears to beconnected to mast cell mediators and propagated by other resident cellswithin target organs or by cells recruited into the sites of mast cellor basophilic degranulation. Corticosteroids which have proven valuablein the management of various allergic diseases and asthma may bebeneficial because of their ability to attenuate this inflammatoryprocess.

Furthermore, there are extra-pulmonary diseases where inflammatoryresponse plays a major role, for example, inflammatory bowel disease,rheumatoid arthritis, glomerulonephritis and inflammatory skin disease.These conditions are also often treated with anti-inflammatory agentswhich may be of short duration or which, like steroidal andnon-steroidal anti-inflammatory drugs, may frequently cause adversesystemic or gastrointestinal reactions.

Improved pharmaceutical treatments for late phase allergic reactions andinflammatory diseases are required.

SUMMARY OF THE INVENTION

It is an object of this invention to provide more effective and safermethods and compositions for treatment of conditions characterized bylate phase allergic reactions or inflammatory reactions.

It is another object of the present invention to provide a method andcompositions for treatment of antigen-induced late phase asthma andbronchial hyperreactivity which do not suffer from the drawbacks of theprior art.

It is a further object of the present invention to provide a method andcompositions for the treatment of asthmatic dual responders which areeffective in preventing and reversing the manifestations of late phaseasthma.

Still another object of the present invention is to provide a method andcompositions as described above which are highly effective indiminishing specific and non-specific bronchial hyperreactivity, andeven when administered after antigen challenge to the patient.

In keeping with these objects and others which will become apparenthereinafter, the invention resides in a method of treating a mammalianpatient suffering from a condition which is characterized by late phaseallergic reactions, including, e.g., pulmonary, nasal, cutaneous, ocularand systemic LPR's, or which is characterized by inflammatory reactions,through the intrabronchial, oral, topical, parenteral, intranasal orintraocular administration to the patient of a pharmaceuticalcomposition comprising from about 0.005 to about 1.0 mg of ultra-lowmolecular weight heparins (ULMWH) per kilogram of patient body weight ineach dose. The administration of these heparins can be on an acute basissuch as following antigen challenge, or on a chronic basis to suppressinflammatory reactions such as bronchial hyperreactivity in asthmapatients.

The ULMWH effective in the method of the invention have averagemolecular weights of about 1,000 to about 3,000 daltons and may exhibita low level of anticoagulant activity or substantially no anticoagulantactivity at all. Novel pharmaceutical compositions are also providedincluding, e.g., inhalant (intrabronchial) compositions in the form ofliquid or powder nebulizer or aerosol compositions containing suitableconcentrations of these ULMWH.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a graph illustrating the effects of antigen challenge on twogroups of allergic sheep, one composed of acute responders and the otherof dual responders. Data for each group are shown as antigen-inducedmean ±SE% change in SR_(L) (specific lung resistance), shown before(baseline), immediately after (P.A.) and up to 8 hours post-antigen.

+=Significantly different from baseline (P<0.05)

*=Significantly different from dual responders (P<0.05)

FIG. 2 comprises two graphs illustrating the differential effects ofinhaled commercial heparin on antigen-induced bronchoconstriction in twogroups of allergic sheep, one composed of acute responders (n=8) and theother of dual responders (n=13).

+=Significantly different from baseline (P<0.05)

*=Significantly different from control (P<0.05)

FIG. 3 is a graph illustrating the effect of pretreatment with inhaledFragmin™ (avg. mol. wt. 5,030 daltons) at 5.0 mg/kg on antigen-inducedbronchoconstriction in dual responder allergic sheep. Data are shown asmean SR_(L) (in cm H₂ O/L/sec) in a group of animals exposed to antigen,first with no drug treatment and again several days later afterpretreatment with Fragmin.

FIG. 4 is a bar graph illustrating the effect of pretreatment withinhaled Fragmin™ at 5.0 mg/kg on AHR in allergic sheep. Data are shownas mean ±SE PD₄₀₀ in breath units at baseline and 24 hours post-antigenchallenge in a group of animals exposed to antigen, first with no drugtreatment and again several days later after pretreatment with Fragmin.

PD₄₀₀ =Cumulative provocating dose of carbachol, increasing SR_(L) to400% above baseline

+=Significantly different from baseline (P<0.05)

FIG. 5 is a graph illustrating the effect of pretreatment with inhaledCY-216 (avg. mol. wt. 4,270 daltons) at 1.25 mg/kg on antigen-inducedbronchoconstriction in dual responder allergic sheep. Data are shown asantigen-induced mean ±SE% change in SR_(L) in a group of animals exposedto antigen, first with no drug treatment and again several days laterafter pretreatment with CY-216.

FIG. 6 is a bar graph illustrating the effect of pretreatment withinhaled CY-216 at 1.25 mg/kg on AHR in allergic sheep. Data are shown asmean ±SE PD₄₀₀ in breath units at baseline and 24 hours post-antigenchallenge in a group of animals exposed to antigen, first with no drugtreatment and again several days later after pretreatment with CY-216.

+=Significantly different from baseline (P<0.05)

FIG. 7 is a graph illustrating the effect of pretreatment with inhaledULMWH CY-222 (avg. mol. wt. 2,355 daltons) at 1.0 mg/kg onantigen-induced bronchoconstriction in dual responder allergic sheep.Data are shown as antigen-induced mean ±SE% change in SR_(L) in a groupof animals exposed to antigen, first with no drug treatment and againseveral days later after pretreatment with CY-222.

*=Significantly different from control (P<0.05)

FIG. 8 is a bar graph illustrating the effect of pretreatment withinhaled CY-222 at 1.0 mg/kg on AHR in allergic sheep. Data are shown asmean ±SE PD₄₀₀ in breath units at baseline and 24 hours post-antigenchallenge in a group of animals exposed to antigen, first with no drugtreatment and again several days later after pretreatment with CY-222.

+=Significantly different from baseline (P<0.05)

FIG. 9 is a graph illustrating the effect of treatment post-antigenchallenge with inhaled CY-222 at 1.0 mg/kg on antigen-inducedbronchoconstriction in dual responder allergic sheep. Data are shown asantigen-induced mean ±SE% change in SR_(L), shown before, immediatelyafter (time zero) and up to 8 hours post-antigen, in a group of animalsexposed to antigen, first with no drug treatment and again several dayslater when CY-222 was administered immediately after the post-antigenmeasurement of SR_(L) (arrow).

*=Significantly different from control (P<0.05)

FIG. 10 is a bar graph illustrating the effect of treatment post-antigenchallenge (arrow in FIG. 9) with inhaled CY-222 at 1.0 mg/kg on AHR inallergic sheep. Data are shown as mean ±SE PD₄₀₀ in breath units atbaseline and 24 hours post-antigen challenge in a group of animalsexposed to antigen, first with no drug treatment and again several dayslater when CY-222 was administered immediately after antigen challenge.

+=Significantly different from baseline (P<0.05)

FIG. 11 is a graph illustrating the effect of pretreatment with inhaledULMWH FRU-70 (avg. mol. wt. 2,500 daltons) at 1.0 mg/kg onantigen-induced bronchoconstriction in dual responder allergic sheep.Data are shown as antigen-induced mean ±SE% change in SR_(L) in a groupof animals exposed to antigen, first with no drug treatment and againseveral days later after pretreatment with FRU-70.

*=Significantly different from control (P<0.05)

FIG. 12 is a bar graph illustrating the effect of pretreatment withinhaled FRU-70 at 1.0 mg/kg on AHR in allergic sheep. Data are shown asmean ±SE PD₄₀₀ in breath units at baseline and 24 hours post-antigenchallenge in a group of animals exposed to antigen, first with no drugtreatment and again several days later after pretreatment with FRU-70.

+=Significantly different from baseline (P<0.05)

FIG. 13 is a graph illustrating the effect of treatment post-antigenchallenge with inhaled FRU-70 at 0.5 mg/kg on antigen-inducedbronchoconstriction in dual responder allergic sheep. Data are shown asantigen-induced mean ±SE% change in SR_(L), shown before, immediatelyafter (time zero) and up to 8 hours post-antigen, in a group of animalsexposed to antigen, first with no drug treatment and again several dayslater when FRU-70 was administered immediately after the post-antigenmeasurement of SR_(L) (arrow) .

*=Significantly different from control (P<0.05)

FIG. 14 is a bar graph illustrating the effect of treatment post-antigenchallenge (arrow in FIG. 13) with inhaled FRU-70 at 0.5 mg/kg on AHR inallergic sheep. Data are shown as mean ±SE PD₄₀₀ in breath units atbaseline and 24 hours post-antigen challenge in a group of animalsexposed to antigen, first with no drug treatment and again several dayslater when FRU-70 was administered immediately after antigen challenge.

+=Significantly different from baseline (P<0.05)

FIG. 15 is a graph illustrating the effect of treatment post-antigenchallenge with inhaled hexasaccharide mixture (avg. mol. wt. 1,930daltons) at 0.5 mg/kg on antigen-induced bronchoconstriction in dualresponder allergic sheep. Data are shown as antigen-induced mean ±SE%change in SR_(L), shown before, immediately after (time zero) and up to8 hours post-antigen, in a group of animals exposed to antigen, firstwith no drug treatment and again several days later when thehexasaccharide mixture was administered immediately after thepost-antigen measurement of SR_(L) (arrow).

*=Significantly different from control (P<0.05)

FIG. 16 is a bar graph illustrating the effect of treatment post-antigenchallenge (arrow in FIG. 15) with inhaled hexasaccharide mixture at 0.5mg/kg on AHR in allergic sheep. Data are shown as mean ±SE PD₄₀₀ inbreath units at baseline and 24 hours post-antigen challenge in a groupof animals exposed to antigen, first with no drug treatment and againseveral days later when the hexasaccharide mixture was administeredimmediately after antigen challenge.

+=Significantly different from baseline (P<0.05)

FIG. 17 is a graph illustrating the effect of treatment post-antigenchallenge with inhaled purified hexasaccharide (avg. mol. wt. 1998daltons) at 0.062 mg/kg on antigen-induced bronchoconstriction in dualresponder allergic sheep. Data are shown as antigen-induced mean ±SE%change in SR_(L), shown before, immediately after (time zero) and up to8 hours post-antigen, in a group of animals exposed to antigen, firstwith no drug treatment and again several days later when thehexasaccharide was administered immediately after the post-antigenmeasurement of SR_(L) (arrow).

*=Significantly different from control (P<0.05)

FIG. 18 is a bar graph illustrating the effect of treatment post-antigenchallenge (arrow in FIG. 17) with inhaled purified hexasaccharide at0.062 mg/kg on AHR in allergic sheep. Data are shown as mean ±SE PD₄₀₀in breath units at baseline and 24 hours post-antigen challenge first,with no drug treatment and again several days later when thehexasaccharide was administered immediately after antigen challenge.

+=Significantly different from baseline (P<0.05)

FIG. 19 is a graph illustrating the effect of treatment post-antigenchallenge with inhaled purified tetrasaccharide (avg. mol. wt. 1290daltons) at 0.062 mg/kg on antigen-induced bronchoconstriction in dualresponder allergic sheep. Data are shown as antigen-induced mean ±SE%change in SR_(L), shown before, immediately after (time zero) and up to8 hours post-antigen, in a group of animals exposed to antigen, firstwith no drug treatment and again several days later when thetetrasaccharide was administered immediately after the post-antigenmeasurement of SR_(L) (arrow).

*=Significantly different from control (P<0.05)

FIG. 20 is a bar graph illustrating the effect of treatment post-antigenchallenge (arrow in FIG. 19) with inhaled purified tetrasaccharide at0.062 mg/kg on AHR in allergic sheep. Data are shown as mean ±SE PD₄₀₀in breath units at baseline and 24 hours post-antigen challenge first,with no drug treatment and again several days later when thetetrasaccharide was administered immediately after antigen challenge.

+=Significantly different from baseline (P<0.05)

FIG. 21 is a graph illustrating the effect of treatment post-antigenchallenge with inhaled octasaccharide (avg. mol. wt. 2,480 daltons) at0.25 mg/kg on antigen-induced bronchoconstriction in dual responderallergic sheep. Data are shown as antigen-induced mean ±SE% change inSR_(L), shown before, immediately after (time zero) and up to 8 hourspost-antigen, in a group of animals exposed to antigen, first with nodrug treatment and again several days later when the octasaccharidemixture was administered immediately after the post-antigen measurementof SR_(L) (arrow).

*=Significantly different from control (P<0.05)

FIG. 22 is a bar graph illustrating the effect of treatment post-antigenchallenge (arrow in FIG. 21) with inhaled octasaccharide at 0.25 mg/kgon AHR in allergic sheep. Data are shown as mean ±SE PD₄₀₀ in breathunits at baseline and 24 hours post-antigen challenge in a group ofanimals exposed to antigen, first with no drug treatment and againseveral days later when the octasaccharide was administered immediatelyafter antigen challenge.

+=Significantly different from baseline (P<0.05)

FIG. 23 is a graph illustrating the effect of pretreatment with inhaleddisaccharide (avg. mol. wt. 660 daltons) at 1.0 mg/kg on antigen-inducedbronchoconstriction in dual responder allergic sheep. Data are shown asantigen-induced mean ±SE% change in SR_(L), shown before, immediatelyafter (time zero) and up to 8 hours post-antigen, in a group of animalsexposed to antigen, first with no drug treatment and again several dayslater after pretreatment with disaccharide.

FIG. 24 is a graph illustrating the effect of pretreatment with orallyadministered purified hexasaccharide (avg. mol. wt. 1,998 daltons) at2.0 mg/kg on antigen-induced bronchoconstriction in a dual responderallergic sheep. Data are shown as antigen-induced an ±SE % change inSR_(L) in a single sheep exposed to antigen, first with no drugtreatment and again several days later after pretreatment withhexasaccharide.

FIG. 25 is a bar graph illustrating the effect of pretreatment withorally administered purified hexasaccharide at 2.0 mg/kg on AHR in anallergic sheep. Data are shown as mean ±SE PD₄₀₀ in breath units atbaseline and 24 hours post-antigen challenge in a single sheep exposedto antigen, first with no drug treatment and again several days laterafter pretreatment with hexasaccharide.

FIG. 26 is a graph illustrating the effect of pretreatment withintravenously administered purified hexasaccharide (avg. mol. wt. 1,998daltons) at 0.25 mg/kg on antigen-induced bronchoconstriction in a dualresponder allergic sheep. Data are shown as antigen-induced mean ±SE%change in SR_(L) in a single sheep exposed to antigen, first with nodrug treatment and again several days later after pretreatment withhexasaccharide.

FIG. 27 is a bar graph illustrating the effect of pretreatment withintravenously administered purified hexasaccharides at 0.25 mg/kg on AHRin allergic sheep. Data are shown as mean ±SE PD₄₀₀ in breath units atbaseline and 24 hours post-antigen challenge in a single sheep exposedto antigen, first with no drug treatment and again several days laterafter pretreatment with hexasaccharide.

FIG. 28 is a bar graph illustrating the comparative activity inpreventing antigen-induced eosinophil influx in the bronchoalveolarlavage fluid of three groups of mice administered, respectively,inhaled, oral and intraperitoneal purified hexasaccharide.

DETAILED DESCRIPTION OF THE INVENTION

The present invention pertains generally to a method of treatment ofmammalian patients suffering from or prone to development of diseaseconditions characterized by late phase allergic reactions and/or byinflammatory reactions, as well as novel pharmaceutical compositionscontaining ultra-low molecular weight heparins which are suitable foruse in practicing said method.

Heparin, a sulfated mucopolysaccharide, is synthesized in mast cells asa proteoglycan and is particularly abundant in the lungs of variousanimals. Heparin is not a specific compound of fixed molecular weightbut is actually a heterogenous mixture of variably sulfatedpolysaccharide chains composed of repeating units of D-glucosamine andeither L-iduronic or D-glucuronic acids. The average molecular weight ofheparin isolated from animal tissues ranges from about 6,000 to about30,000 daltons.

Pharmacologically, heparin is known primarily as an anticoagulant. Thisactivity results from heparin's ability to bind to some of the residuesof antithrombin III (AT-III), accelerating the neutralization by AT-IIIof activated clotting factors and preventing the conversion ofprothrombin to thrombin. Larger amounts of heparin can inactivatethrombin and earlier clotting factors, preventing conversion offibrinogen to fibrin.

The hemorrhagic activity of heparin is related to the molecular weightof its polysaccharide fragments; low molecular weight components orfragments (for example, fragments having a molecular weight of less than6,000 daltons) have moderate to low antithrombin and hemorrhagiceffects. Similarly, low molecular weight heparins isolated from animaltissue generally have reduced hemorraghic properties compared tocommercial heparin but may still have significant anticoagulantactivity.

Commercial heparin, which is generally derived from beef lung or porkintestinal mucosa, has an average molecular weight of about15,000-17,500 daltons.

Heparin has been shown to act as a specific blocker of the IP₃ receptorsand inhibits IP₃ mediated calcium release. We have previously suggestedthat heparin may block IP₃ receptors in mast cells and therefore byinterfering with signal transduction may modulate mast celldegranulation and mediator release. In vivo and in vitro studies supportthis concept and have demonstrated that inhaled heparin can attenuateallergic bronchoconstriction in sheep, prevent exercise induced asthma,as well as inhibit anti IgE induced mast cell histamine release. Inhaledheparin in doses up to 1,000 units/kg has been found to have no effecton partial thromboplastin time (PTT), thus, suggesting a"non-anticoagulant" effect.

It has also been reported that low molecular weight heparins (averagemolecular weight about 4,500 daltons), which have reduced APTT activity,were effective in animal studies in preventing antigen-inducedbronchoconstrictor response (ABR) and bronchial hyperreactivity, alsoreferred to as airway-hyperresponsiveness (AHR). However, as discussedand illustrated in greater detail below, neither commercial heparin normedium or low molecular weight heparins, even those with very lowanticoagulant activity, are effective in ameliorating AHR subsequent toantigen challenge in test animals. These heparins apparently provideonly a prophylactic, preventive effect, but are not of value in treatingan antigen-triggered asthmatic episode.

We have discovered and reported in parent application Ser. No.08/516,786 that ultra-low molecular weight heparin (ULMWH) fractions arenot only effective inhibitors of airway anaphylaxis, but are highlyeffective in reducing AHR even when administered after antigenchallenge. Chronic, regular use of ULMWH may also reduce AHR, and ULMWHtherefore may be used for chronic therapy of asthma whether caused byspecific (i.e., antigenic) or non-specific factors.

Our prior application only pertained to, and disclosed test datademonstrating, the efficacy of ULMWH in the treatment of early phaseasthma in acute responders, not in the treatment of "dual responders"who experience both early and late phase bronchoconstriction andprolonged AHR. As discussed previously, it could not have been predictedbased on our earlier studies that ULMWH, whether administered before orafter antigen challenge, would be efficacious in inhibitingbronchoconstriction (both early and late phase) and AHR in dualresponders. This lack of predictability is evidenced by the fact thatcommercial heparin and medium or low molecular weight heparins (mol.wt. >3,000) inhibit AHR in acute responders when administered beforeantigen challenge, but have no significant effect in suppressing thelate phase reaction and AHR observed in dual responders.

After conducting further controlled studies with ULMWH, we discovered,surprisingly, that heparin fractions having average molecular weights ofabout 1,000 to about 3,000 are effective, when inhaled by dualresponders prior to or even after antigen challenge, in suppressingearly and late phase bronchoconstriction and AHR.

Even more surprisingly, we found that oral and intravenous (or otherparenteral) administration of ULMWH prior to antigen challengeeffectively inhibited bronchoconstriction and AHR in dual responders.

Accordingly, the present invention comprises in one aspect a method oftreating a mammalian patient who is a dual responder and suffers fromantigen-induced late phase asthma comprising the intrabronchialadministration to the patient before or after antigen challenge of apharmaceutical composition containing about 0.005 to about 1.0 mg of oneor more effective ULMWH fractions per kilogram of patient body weight ineach dose of said composition, and preferably from about 0.075 to about0.75 mg/kg per dose. For purposes of this application, the "effectiveULMWH" may be defined as heparin fractions having an average molecularweight of about 1,000-3,000 daltons. ULMWH having an average molecularweight of about 1,000-2,500 daltons are particularly effective when usedin the method of the invention. Each ULMWH fraction may comprisetetrasaccharides, pentasaccharides, hexasaccharides, septasaccharides,octasaccharides and decasaccharides as well as molecules of greaterchain length.

The ULMWH fractions used in the invention are oligomers of sulfatedsaccharide units which may have, e.g., the following general structuralformula: ##STR1##

Despite the known activity of N-desulfated heparins in other biologicalsystems, for example as inhibitors of cell growth, it has been foundthat the saccharide units in the ULMWH fractions which are effective forpurposes of the present invention are all N-sulfated; N-desulfatedfractions are ineffective.

While the sulfated polysaccharides used in the method and compositionsof the invention are generally referred to herein as ultra-low molecularweight heparins, i.e., ultra-low molecular weight fractions derived fromnaturally occurring heparin (or synthetic versions of such ULMWH), theinvention may also encompass the use of sulfated polysaccharides derivedfrom heparan sulfate, dermatan sulfate, chondroitin sulfate, pentosanpolysulfate and/or other glycosaminoglycans and mucopolysaccharides. Thesubject sulfated polysaccharide fractions must, however, have an averagemolecular weight of about 1,000-3,000 daltons. Pharmaceuticallyacceptable salts of the effective ULMWH or any of the other sulfatedpolysaccharides listed above may also be utilized, e.g., the sodium,calcium or potassium salts.

In accordance with this first aspect of the invention, a human or othermammalian patient who is a dual responder who has inhaled, ingested orotherwise come into contact with an antigen (i.e., has been "challenged"with an antigen) of a type known to provoke asthmatic episodes in thatpatient, or a patient who may be exposed at a future time to antigenchallenge, is administered via inhalation at least one dose of apharmaceutical composition containing one or more effective ULMWHcumulatively present in the above-described concentration ranges.Additional doses may subsequently be administered as necessary afterantigen challenge until the patient regains or maintains normal airflowresistance levels.

The invention also comprehends in a second aspect the chronicadministration of effective ULMWH to dual responder asthma patients toreduce and suppress early and late phase AHR. "Chronic administration"as used herein refers to administration of pharmaceutical compositionscontaining effective ULMWH at least once daily for at least tenconsecutive days. Chronic administration of a composition containingfrom about 0.005-1.0 mg/kg per dose, and preferably about 0.0075-0.75mg/kg per dose, can be continued indefinitely to provide AHR-suppressanttherapy at least comparable to corticosteroids but substantially withoutside effects.

The inhalant (intrabronchial) ULMWH compositions used in the presentinvention to treat late phase asthma and other pulmonary conditions maycomprise liquid or powder compositions containing effective ULMWHfractions and suitable for nebulization and intrabronchial use, oraerosol compositions administered via an aerosol unit dispensing metereddoses.

Suitable liquid compositions comprise for example, effective ULMWH in anaqueous, pharmaceutically acceptable inhalant solvent, e.g., isotonicsaline or bacteriostatic water. The solutions are administered by meansof a pump or squeeze-actuated nebulized spray dispenser, or by any otherconventional means for causing or enabling the requisite dosage amountof the liquid composition to be inhaled into the mammalian patient'slungs.

Suitable powder compositions include, by way of illustration, powderedpreparations of heparin thoroughly intermixed with lactose or otherinert powders acceptable for intrabronchial administration. The powdercompositions can be administered via an aerosol dispenser or encased ina breakable capsule which may be inserted by the mammalian patient intoa device that punctures the capsule and blows the powder out in a steadystream suitable for inhalation.

Aerosol formulations for use in the subject method would typicallyinclude fluorinated alkane propellants, surfactants and co-solvents andmay be filled into aluminum or other conventional aerosol containerswhich are then closed by a suitable metering valve and pressurized withpropellant, producing a metered dose inhaler (MDI).

The total concentration of effective ULMWH in any propellant vehiclesuitable for use in a pressured aerosol dispenser, such as an MDI, mustbe sufficiently high to provide a dose of about 0.005-0.1 mg (5-100 μg)of effective ULMWH per kilogram of patient body weight peradministration. Thus, for example, if an MDI delivers about 85 μl ofdrug-containing propellant vehicle per actuation, the concentration ofeffective ULMWH in the vehicle in the case of a mammalian patientweighing 75 kg would be approximately 0.0045-0.088 mg/μl (4.5-88 μg/μl),delivering 0.375 to 7.5 mg (375-7,500 μg) of ULMWH per actuation, if itis desired to deliver the entire dose with a single actuation. If atwo-actuation dose is desired, the corresponding concentration rangewould be approximately 0.0022-0.044 mg/μl (2.2-44 μg/μp), delivering0.188 to 3.75 mg (188-3,750 μg) of ULMWH per actuation.

The total concentration of effective ULMWH in any liquid nebulizersolution must be sufficiently high to provide a dose of about 0.05-1.0mg (50-1000 μg) of effective ULMWH per kilogram of patient body weightper administration. Thus, for example, if the nebulizer utilizeddelivers 5 ml of solution per actuation, the concentration of effectiveULMWH in the case of a mammalian patient weighing 75 kg should beapproximately 0.75-15.0 mg/ml.

In a further aspect of the invention, effective ULMWH-containingcompositions are administered orally or parenterally (e.g., IV or IM) tomammalian patients suffering from antigen-induced late phase asthma,i.e., who are dual responders, prior to exposure of the patient toantigen-challenge. The oral or parenteral compositions contain about0.005 to about 1.0 mg of effective ULMWH per kg of patient body weightin each dose. The oral or parenteral compositions may be administered upto 8 hours (but preferably not more than 4 hours) prior to antigenchallenge and are effective in reducing early and late phasebronchoconstriction and in suppressing AHR.

As those skilled in the pharmaceutical arts will appreciate, manyconventional methods and apparatus are available for administeringprecisely metered doses of intrabronchial medicaments and for regulatingthe desired dosage amount in accordance with patient weight and theseverity of the patient's condition. Moreover, there are manyart-recognized liquid, powder and aerosol vehicles suitable for theintrabronchial ULMWH compositions of the present invention, and manypharmaceutically acceptable oral and parenteral vehicles which may beemployed for the oral and parenteral ULMWH-containing compositions. Theinvention is not limited to any particular inert vehicles, solvents,carriers excipients or dosage forms and is not restricted to anyparticular methods or apparatus of intrabronchial administration.

The pharmaceutical compositions may also be dosage forms which containthe effective ULMWH as active ingredients in any pharmaceuticallyacceptable oral, injectable or IV dosage vehicles, or in topical orintraocular vehicles. Each dosage form includes about 0.005-1.0 mg/kg ofaverage patient body weight of effective ULMWH (one or a combination ofULMWH) and pharmaceutically acceptable inert ingredients, e.g.,conventional excipients, vehicles, fillers, binders, disintegrants,solvents, solubilizing agents, sweeteners, coloring agents and any otherinactive ingredients which are regularly included in pharmaceuticaldosage forms for oral administration. Suitable oral dosage forms includetablets, capsules, caplets, gelcaps, pills, liquid solutions,suspensions or elixirs, powders, lozenges, micronized particles andosmotic delivery systems. Injectable and IV dosage forms includeisotonic saline solutions or dextrose solutions containing suitablebuffers and preservatives. Many suitable dosage forms and vehicles, andlistings of inactive ingredients therefor, are well-known in the art andare set forth in standard texts such as Remington's PharmaceuticalSciences, 17th edition (1985).

The ULMWH compositions described herein provide highly effectivetreatment for early and late phase antigen-induced asthma even afterantigen challenge has occurred, as well as for other conditionscharacterized by late phase allergic reactions.

To demonstrate the unexpected superiority of the effective ULMWH incomparison with higher molecular weight heparins in treating asthmaticdual responders, experiments were conducted comparing the effects ofdifferent heparin types on dual responder allergic sheep, both beforeand after antigen challenge. Detailed descriptions of these experimentsand of the results obtained are set forth in the following examples aswell as in the graphs shown in the drawings.

The following examples, while illustrating the methods and compositionsof the invention and demonstrating the efficacy of the same, are notintended to set forth specific compositions, materials, procedures ordosage regimens which must be utilized exclusively in order to practicethe invention.

EXAMPLE 1 Administration of Inhaled ULMWH to Dual Responder AllergicSheep Methods

Pulmonary Airflow Resistance:

Allergic sheep with previously documented dual bronchoconstrictorresponse to Ascaris suum antigen were used for all studies. The sheepwere intubated with a cuffed nasotracheal tube and pulmonary airflowresistance (R₁) was measured by the esophageal balloon cathetertechnique, while thoracic gas volume was measured by bodyplethysmography. Data were expressed as specific R_(L) (SR_(L), definedas R_(L) times thoracic gas volume (V_(tg))).

Airway Responsiveness:

To assess airway responsiveness, cumulative dose-response curves toinhaled cabachol were performed by measuring SR_(L) before and afterinhalation of buffered saline and after each administration of 10breaths of increasing concentrations of carbachol (0.25, 0.5, 1.0, 2.0and 4.0% wt/vol solution). Airway responsiveness was measured bydetermining the cumulative provocation dose (PD₄₀₀) of carbachol (inbreath units) that increased SR_(L) to 400% above baseline. One breathunit was defined as one breath of 1% carbachol solution.

Heparin Fractions:

In the studies reported herein various heparin materials wereadministered to the dual responder allergic sheep prior to and/or afterantigen challenge. Some of these heparins were ULMWH fractions ofaverage molecular weight between 1,000 and 3,000 daltons, some were ofhigher average molecular weight, and one was of lower molecular weight.The heparin fractions tested are set forth in Table 1 below.

                  TABLE 1                                                         ______________________________________                                        HEPARIN FRACTIONS AND THEIR MOLECULAR WEIGHTS                                     FRACTION        AVG. MOL. WT.                                                                              CATEGORY                                     ______________________________________                                        Commercial heparin                                                                            15,000 d     Heparin                                            Fragmin ™   5,030 d Low mol. wt.                                           CY-216  4,270 d Low mol. wt.                                                  CY-222.sup.1 (Sanofi)  2,355 d ULMWH                                          FRU-70.sup.2 (Kabivitrum)  2,500 d ULMWH                                      Hexasaccharide mixture.sup.3  1,930 d ULMWH                                   Octasaccharide.sup.4  2,480 d ULMWH                                           Purified hexasaccharide.sup.5  1,998 d ULMWH                                  Purified tetrasaccharide.sup.6  1,290 d ULMWH                                 Disaccharide.sup.7   660 d Sub-ULMWH                                        ______________________________________                                         .sup.1 An anticoagulant octasaccharide mixture.                               .sup.2 A nonanticoagulant octasaccharide mixture.                             .sup.3 Oligosaccharide derived from commercial porcine heparin, comprisin     primarily tetrasaccharide, hexasaccharide, octasaccharide and                 decasaccharide fractions.                                                     .sup.4 Obtained from the hexasaccharide mixture by gel column                 chromatography, comprises about 70% octasaccharide and 30% decasaccharide     fractions.                                                                    .sup.5 Obtained from the hexasaccharide mixture by gel column                 chromatography.                                                               .sup.6 Obtained from the hexasaccharide mixture by gel column                 chromatography.                                                               .sup.7 The disaccharide was trisulfated but had a molecular weight so low     it could not be considered a ULMWH fraction with heparinlike properties. 

Experimental Protocol

Airway Studies

Each animal's baseline airway responsiveness (PD₄₀₀) was determined, andthen on different experimental days the sheep underwent airway challengewith Ascaris suum antigen. SR_(L) was measured, before and immediatelyafter challenge, and then hourly for 8 hours. The post-challenge PD₄₀₀was measured 24 hours after antigen challenge when AHR occurred. Theprotocol was repeated at least 14 days later, but each animal wasadministered a dose of one of the test heparin fractions either about 30minutes before antigen challenge or immediately after post-challengeSR_(L) measurement.

Data Analysis

Data were expressed as: ##EQU1##

Results

FIG. 1 illustrates the differential reactions to antigen challenge oftwo groups of allergic sheep, one composed of acute responders and theother of dual responders. The SR_(L) of the acute responders returned toapproximately baseline levels after about three hours post-antigen andremained there. In the dual responders, however, there is a late phasepeak in SR_(L) at about six hours with levels remaining significantlyabove baseline through the eight-hour end point of the study. It is thissecond, late phase peak which characterizes dual responders.

FIGS. 2A and 2B depict the effects of pre-challenge treatment withinhaled commercial heparin on SR_(L) in acute responders (FIG. 2A) andin dual responders (FIG. 2B). While the SR_(L) of the acute respondersremained at near-baseline levels even after antigen challenge, both theearly phase and the late phase SR_(L) in the dual responders was notalleviated by heparin pretreatment, even in dual responders administeredas much as 2000 units per kilogram.

FIGS. 3-6 illustrate the lack of efficacy of the low molecular weightheparin fractions, Fragmin and CY-216, in modifying eitherbronchoconstriction or AHR in dual responders when administered prior toantigen challenge.

FIGS. 7-10 show that both pretreatment and post-antigen challengetreatment with the inhaled ULMWH CY-222 (avg. mol. wt. 2355 d, withinthe range of effective ULMWH in accordance with the invention) wereeffective in significantly modifying both early and late phaseantigen-induced bronchoconstriction and AHR in the dual responders.

FIGS. 11-14 illustrate the efficacy of both pretreatment andpost-antigen challenge treatment with the ULMWH FRU-70 (avg. mol. wt.2500 d) in the treatment of early and late phase asthma.

FIGS. 15-22 demonstrate the efficacy of various effective ULMWHfractions, even when administered post-antigen challenge, insignificantly reducing bronchoconstriction and AHR in dual responders.

FIG. 23 shows that the disaccharide fraction, having an averagemolecular weight of only about 660 d (substantially below the weightrange required for the effective ULMWH fractions) was ineffective inmodifying antigen-induced bronchoconstriction in the dual responderallergic sheep.

In the experiments whose data are reflected in FIGS. 7-14 and 17-22 thedosage of the effective ULMWH administered to the allergic sheep was thelowest effective dose (determined through dose-ranging trials) for eachULMWH fraction. It will be observed that the different ULMWH had varyingminimum effective dose levels in the treatment of dual responders. Theminimum effective dose was about 1.0 mg/kg for CY-222 and for FRU-70administered prior to antigen challenge, but about 0.5 mg/kg for FRU-70administered post-antigen challenge and for the inhaled hexasaccharidemixture. The purified tetrasaccharide, having the lowest averagemolecular weight of any of the effective ULMWH tested, had a minimumeffective dose when administered post-antigen of 0.062 mg/kg, as did thepurified hexasaccharide. These data tend to suggest that purifiedfractions having an average weight near the lower limit of about 1,000 dmay be the most effective ULMWH, at least in the treatment of dualresponders. The optimal structural domain and/or sequence for theobserved antiallergic and/or anti-inflammatory activity appears to bethe tetrasaccharide.

EXAMPLE 2 Administration of Oral ULMWH to Dual Responder Allergic Sheep

The procedure of Example 1, in terms of the test animals and evaluationmethods, were followed in the present experiment.

One dual-responder allergic sheep was administered orally 2 mg/kg ofpurified hexasaccharide (avg. mol. wt. 1998 daltons) 90 minutes beforechallenge with Ascaris Suum antigen. The effects of the pretreatmentwith the hexasaccharide on SR_(L) from baseline (time of administrationof the hexasaccharide) through 8 hours after antigen challenge isreflected on FIG. 24. Also shown on FIG. 24 for comparison purposes isthe percentage change in SR_(L) in the same dual responder sheep (in anexperiment conducted several days earlier) challenged with antigen butwithout ULMWH pretreatment.

Shown in FIG. 25 are the respective PD₄₀₀ values measured at baselineand post-antigen when the sheep was administered antigen challenge withhexasaccharide pretreatment and with no pretreatment (control).

EXAMPLE 3 Administration of Intravenous ULMWH to Dual Responder AllergicSheep

The procedure of Example 2 was followed with another dual responderallergic sheep, except that 0.25 mg/kg of purified hexasaccharide wasadministered intravenously one hour before antigen challenge in oneexperiment, while antigen was administered with no pretreatment in thesecond (control) experiment. The percentage change in SR_(L) for thepretreatment and control experiments are shown in FIG. 26 and the PD₄₀₀values for those experiments at baseline and post-antigen are shown inFIG. 27.

EXAMPLE 4 Prevention of Antigen-induced Eosinophil Influx in Mice

In four groups of sensitized laboratory mice (n=3 in each group)bronchoalveolar lavage was performed 24 hours after antigen challenge todetermine eosinophil influx values in each group. The mice were treatedwith either aerosolized saline (placebo) or purified hexasaccharideadministered by the following routes and in the following dosageamounts, respectively: inhaled aerosol,⁸ oral (100 μg) andintraperitoneal (40 μg). The percentage inhibition of eosinophil influxeffected in each treatment group was determined by comparing the levelof such influx measured in bronchoalveolar ravage fluid subsequent tohexasaccharide administration with the saline group.

The mean percentage inhibition values for the three treatment groups ofmice are reflected in FIG. 28. The mice receiving inhaled and oralhexasaccharide showed a 40-50% reduction in eosinophil influx while themice receiving intraperitoneal hexasaccharide showed about a 20%reduction in such influx.

⁸ The mice (n=3) were placed in a chamber containing 10 mg ofhexasaccharide in 9 ml of bacteriostatic injection water, which wasaerosolized. The mice were allowed to inhale the aerosol for about 30minutes.

The differential effects of commercial heparin observed in acute anddual responders (shown in FIGS. 2A and 2B) might indicate theinvolvement of different signaling pathways during airway anaphylaxis.This would suggest that during immunologically mediated mast-cellreaction in the airways, IP₃ is the predominantly active pathway in"acute responders" while non-IP₃ pathways (e.g., diacyl-glycerol/proteinkinase C or other pathways) may be operative in "dual responders".

The late phase response and AHR are associated with marked airwayinflammation. The pathological studies of the airway mucosa andbronchoalveolar lavage (BAL) have shown influx of eosinophils,neutrophils and activated T-lymphocytes during this phase. Increasedlevels of eosinophil-derived inflammatory mediators in plasma and BAL,including eosinophilic cationic protein and major basic protein, havebeen observed during the late phase reaction. Upregulation of TH₂ -typecytokines (IL₄ and IL₅) following allergen challenge has also beenobserved during the late phase. Thus, the cellular inflammatoryresponse, in combination with released pro-inflammatory mediators (e.g.,leukotrienes, PAF, eosinophilic proteins, etc.) and locally producedcytokines in the bronchial mucosa, play a central role in the late phaseallergic inflammation and bronchoconstriction.

AHR and thus airway inflammation can be modified either by prevention ofmast mediator release by "anti-allergic agents" (e.g., cromolyn sodium)or by the action of "anti-inflammatory" agents likeglucocorticosteroids. The "anti-allergic" agents are only effective asprophylactic agents and can prevent the mediator release and AHR.Because these agents do not possess anti-inflammatory activity, they aregenerally ineffective when administered after the exposure to antigen.By contrast "anti-inflammatory" agents can attenuate post-antigen AHRand airway inflammation, whether administered before or after theexposure to antigen. Our data suggest that the actions of ULMWH areanalogous to the anti-inflammatory actions of glucocorticosteroids.

Since the effective ULMWH can modify AHR even when administered afterantigen challenge, they should also be useful in the treatment ofnon-asthmatic conditions associated with AHR, e.g., chronic bronchitis,emphysema and cystic fibrosis.

Moreover, in view of our findings regarding the efficacy of certainULMWH in inhibiting asthmatic LPR in a manner resembling theanti-inflammatory effects of corticosteroids, the effective ULMWH shouldbe useful in the treatment of the following conditions and by thefollowing routes of administration:

1. Late phase reactions and inflammatory response in extra-pulmonarysites:

(a) allergic rhinitis

(b) allergic dermatitis

(c) allergic conjunctivitis

2. Extra-pulmonary diseases where inflammatory response plays a majorrole:

(I) inflammatory bowel disease

(ii) rheumatoid arthritis and other collagen vascular diseases

(iii) glomerulonephritis

(iv) inflammatory skin diseases

(v) sarcoidosis

3. Routes of Administration

(I) intrabronchial

(ii) intranasal

(iii) intraocular

(iv) topical

(v) oral

(vi) parenteral (IM or IV)

It should be emphasized, however, that the invention is not restrictedor limited in any way to any theoretical or actual physiological orbiochemical mechanisms or pathways, but comprehends the methods oftreatment of conditions characterized by late phase allergic reactions,or treatment of dual responder mammalian patients, and the compositionsfor use in said methods described hereinabove, notwithstanding theactual mechanisms of action involved.

It has thus been shown that there have been provided methods andcompositions which achieve the various objects of the invention andwhich are well adapted to meet the conditions of practical use.

As various possible embodiments might be made of the above invention,and as various changes might be made in the embodiments set forth above,it is to be understood that all matters herein described are to beinterpreted as illustrative and not in a limiting sense.

What is claimed as new and desired to be protected by letters patent isset forth in the following claims.

I claim:
 1. A method of treating a mammalian patient suffering from orprone to a condition whose symptoms include late phase allergicreactions, airway hyperresponsiveness or inflammatory reactions, saidmethod comprising the administration to the patient of a pharmaceuticalcomposition containing about 0.005 to about 1.0 mg of ultra-lowmolecular weight heparins (ULMWH) per kilogram of patient body weight ineach dose, said ULMWH having an average molecular weight of about 1,000to about 3,000 daltons.
 2. A method according to claim 1 wherein saidULMWH have an average molecular weight of about 1,000 to about 2,500daltons.
 3. A method according to claim 1 wherein said ULMWH compriseheparin fractions selected from the group consisting oftetrasaccharides, pentasaccharides, hexasaccharides, septasaccharides,octasaccharides and decasaccharides and pharmaceutically acceptablesalts thereof.
 4. A method according to claim 1 wherein said ULMWH areN-sulfated.
 5. A method according to claim 1 wherein said compositioncontains about 0.075 to about 0.75 mg of said ULMWH per kilogram perdose.
 6. A method according to claim 1 wherein said ULMWH arenon-anticoagulant.
 7. A method according to claim 1 wherein saidcomposition is administered by the oral, parenteral, topical,intrabronchial, intranasal or intraocular routes.
 8. A method accordingto claim 7 wherein said parenteral administration is intravenous orintramuscular.
 9. A method according to claim 1 wherein said symptomsare late phase allergic reactions selected from the group consisting ofpulmonary late phase reactions, nasal late phase reactions, cutaneouslate phase reactions, ocular late phase reactions and systemic latephase reactions.
 10. A method according to claim 9 wherein said latephase reactions are pulmonary late phase reactions.
 11. A methodaccording to claim 10 wherein said condition is late phase asthma.
 12. Amethod according to claim 1 wherein said condition is a non-asthmaticcondition characterized by airway hyperresponsiveness.
 13. A methodaccording to claim 12 wherein said condition is selected from the groupconsisting of chronic bronchitis, emphysema and cystic fibrosis.
 14. Amethod according to claim 1 wherein said symptoms are inflammatoryreactions.
 15. A method according to claim 1 wherein said condition isselected from the group consisting of allergic rhinitis, allergicdermatitis, allergic conjunctivitis, inflammatory bowel disease,rheumatoid arthritis, collagen vascular diseases, glomerulonephritis,inflammatory skin diseases and sarcoidosis.
 16. A method according toclaim 1 wherein said composition comprises a solution or suspension ofsaid ULMWH in an aqueous, pharmaceutically acceptable, liquid inhalantvehicle.
 17. A method according to claim 16 wherein said vehicle isisotonic saline or bacteriostatic water.
 18. A method according to claim16 wherein said composition is administered by means of a pump orsqueeze-actuated nebulizer.
 19. A method according to claim 16 wherein asufficient amount of said composition is administered to the patient toprovide a dose of about 0.05-1.0 mg/kg of said ULMWH.
 20. A methodaccording to claim 16 wherein said composition contains about 0.75-15.0mg/ml of said ULMWH.
 21. A method according to claim 1 wherein saidcomposition is an aerosol composition comprising an aerosol propellant.22. A method according to claim 21 wherein said composition is dispensedvia a metered dose inhaler.
 23. A method according to claim 21 wherein asufficient amount of said composition is administered to the patient toprovide a dose of about 0.005-0.1 mg/kg of said ULMWH.
 24. A methodaccording to claim 21 wherein said composition contains about 2.2-88μg/μl of said ULMWH.
 25. A method according to claim 1 wherein saidcomposition comprises a powdered preparation of said ULMWH intermixedwith an inert powder acceptable for intrabronchial administration.
 26. Amethod according to claim 25 wherein said inert powder is lactose.
 27. Amethod according to claim 25 wherein said composition is administeredvia an aerosol dispenser.
 28. A method according to claim 25 whereinsaid composition is administered from a breakable capsule.
 29. A methodaccording claim 1 wherein said composition is administered to thepatient prior to challenge with allergic reaction-inducing antigen. 30.A method according claim 1 wherein said composition is administered tothe patient subsequent to challenge with allergic reaction-inducingantigen.
 31. A method of treating a mammalian patient suffering from orprone to a condition whose symptoms include late phase allergicreactions, airway hyperresponsiveness or inflammatory reactions, saidmethod comprising the administration to the patient of a pharmaceuticalcomposition containing about 0.005 to about 1.0 mg of sulfatedpolysaccharides per kilogram of patient body weight in each dose, saidsulfated polysaccharides having an average molecular weight of about1,000 to about 3,000 daltons.
 32. A method according to claim 31 whereinsaid sulfated polysaccharides are derived from glycosaminoglycans ormucopolysaccharides.
 33. A method according to claim 32 wherein saidsulfated polysaccharides are derived from heparin, heparan sulfate,dermatan sulfate, chondroitin sulfate or pentosan polysulfate.
 34. Amethod according to claim 31 wherein said sulfated polysaccharidescomprise tetrasaccharides, pentasaccharides, hexasaccharides,septasaccharides, octasaccharides and decasaccharides andpharmaceutically acceptable salts thereof.
 35. A method according toclaim 34 wherein said sulfated polysaccharides comprisetetrasaccharides.